Abstract: The present application has a proposition to provide a highly efficient laser excitation fluorescent microscope. Accordingly, a laser excitation fluorescent microscope of the present application includes a laser light source part radiating at least two types of excitation lights having different wavelengths; a light collecting part collecting the two types of excitation lights on a sample; a high-functional dichroic mirror, disposed between the laser light source part and the light collecting part, reflecting the two types of excitation lights to make the excitation lights incident on the light collecting part, and transmitting two types of fluorescence generated at the sample; and a detecting part detecting light transmitted through the high-functional dichroic mirror, in which an incident angle ? of the excitation lights and the fluorescence to the high-functional dichroic mirror satisfies a formula of 0°<?<45°.

Abstract: The present application generates an image suitable for collectively observing a whole area or all wavelength components in a viewing field. A confocal microscope apparatus includes a detecting unit being disposed on a collecting location of the collecting optical system, separating incident light into a light from a vicinity of a collecting point on the sample and a light from a peripheral of the vicinity, and detecting each of the lights; and an image generating unit generating an image of the sample by a light signal from the vicinity and a light signal from the peripheral of the vicinity, and setting a ratio of the signal of the light from the vicinity of the collecting point to the signal of the light from the peripheral of the vicinity for each of areas on the image.

Abstract: A confocal microscope apparatus is capable of obtaining a high confocal effect while detecting an image of a specimen line by line. The confocal microscope apparatus may include an illuminating optical system which illuminates a line-shaped area on a specimen plane in a specimen by collected light, an image-forming optical system which forms an image of light emitted from the specimen plane, a two-dimensional light detector which is placed at a conjugate plane of the specimen plane, a scanning unit which moves the line-shaped area on the specimen plane, and a correcting unit which corrects a pixel signal of a specific line on the two-dimensional light detector having a confocal relation with the line-shaped area based on a pixel signal of a peripheral line of the specific line.

Abstract: A spectral image processing method is capable of reducing noise while maintaining necessary information. The spectral image processing method performs processing on a spectral image of a specimen, including a step of normalizing spectra (=spectral brightness curves) of respective pixels constituting the spectral image such that their brightness levels become equal, a step of smoothing the normalized spectra in spatial directions of the respective pixels, and a step of denormalization of multiplying spectra of the respective pixels obtained by the smoothing by either one of brightness levels of the pixels corresponding the spectra and values corresponding to the brightness levels. Consequently, the noise can be reduced while information on brightness distribution on the image is maintained.

Abstract: A spectral image processing system and method of performing robust unmixing on measurement noise. Based on an observed spectral image acquired from a specimen and emission spectral data of each of plural materials contained in the specimen, a contribution of each of the plural materials to the observed spectral image is unmixed by a process, including an evaluating step of evaluating reliability of each component of the observed spectral image based on a predicted spectral image of the observed spectral image, and a reflecting step of reflecting a result of the evaluation in a content of the unmixing.

Abstract: The present application has a proposition to provide a highly efficient laser excitation fluorescent microscope. Accordingly, a laser excitation fluorescent microscope of the present application includes a laser light source part radiating at least two types of excitation lights having different wavelengths; a light collecting part collecting the two types of excitation lights on a sample; a high-functional dichroic mirror, disposed between the laser light source part and the light collecting part, reflecting the two types of excitation lights to make the excitation lights incident on the light collecting part, and transmitting two types of fluorescence generated at the sample; and a detecting part detecting light transmitted through the high-functional dichroic mirror, in which an incident angle ? of the excitation lights and the fluorescence to the high-functional dichroic mirror satisfies a formula of 0°<?<45°.

Abstract: A proposition of the present invention is to acquire necessary data without any chasm in an observation of a specimen containing plural kinds of substances of which exciting wavelengths are different. Accordingly, a spectral observation method of the present invention, irradiating light to the specimen containing plural kinds of substances (RFP, GFP, and so on) of which exciting wavelengths are different, and detecting spectra of light emitted from the specimen, the spectral observation method sequentially acquires spectral data of substances from the one of which exciting wavelength is long while switching a wavelength of the irradiated light among respective exciting wavelengths of the plural kinds of substances, and excludes the exciting wavelength of the substance corresponding to the spectral data to be acquired from detection wavelengths of the spectra each when acquiring the spectral data of the spectral data of the respective substances.

Abstract: It is an object to realize smooth switching between free scanning and high speed scanning in a light scanning apparatus and a light scanning microscope. To attain the object, a light scanning apparatus includes at least three mirror scanners disposed at predetermined positions of a light path for light scanning, and a light path switching unit switching the light path between a light path in which a highest-speed mirror scanner among the mirror scanners is valid and a light path in which the highest-speed mirror scanner is invalid. Therefore, the switching between a free scanning mode and a high speed scanning mode is performed by the driving of the light switching unit and involves no movement of galvanometer scanners. In addition, since the resonant galvanometer scanner is invalid during the free scanning mode, it is possible to make the resonant galvanometer scanner on standby.

Abstract: The present application generates an image suitable for collectively observing a whole area or all wavelength components in a viewing field. A confocal microscope apparatus includes a detecting unit being disposed on a collecting location of the collecting optical system, separating incident light into a light from a vicinity of a collecting point on the sample and a light from a peripheral of the vicinity, and detecting each of the lights; and an image generating unit generating an image of the sample by a light signal from the vicinity and a light signal from the peripheral of the vicinity, and setting a ratio of the signal of the light from the vicinity of the collecting point to the signal of the light from the peripheral of the vicinity for each of areas on the image.

Abstract: The present application detects a state change of an object in an optical axis direction by a single frame scanning for each round. A confocal microscope apparatus includes a detecting unit separating incident light into a light from a vicinity of a collecting point on the sample and a light from a peripheral of the vicinity and detecting each of the lights, and an image generating unit generating an image of the sample by a light signal from the vicinity and a light signal from the peripheral, in which a distinction between a light signal from the vicinity and a light signal from the peripheral is calculated for each of a plurality of images generated in a number of times of scanning without changing a collecting location in an optical axis direction.

Abstract: To provide a laser scanning microscope capable of enhancing the degree of freedom of observation while keeping its structure simple. Accordingly, a laser scanning microscope includes a light source, a spectroscopic unit guiding light from the light source to a specimen and guiding the light from the specimen to a detector, light path switching units switching a light path between the spectroscopic unit and the specimen to one among a plurality of light paths with different routes, and a plurality of light deflecting units each disposed in each of the plurality of light paths.

Abstract: A microscope system includes a casing in which an optical element is housed, an airtight specimen chamber, and an environment control unit that controls temperature and humidity in the specimen chamber and also controls temperature inside the casing. An upper plate of the specimen chamber is an open/close lid opened and closed when inserting or removing a biological specimen. The casing becomes sealed by a bottom plate of the specimen chamber as the specimen chamber is set on the casing.

Abstract: To provide a laser scanning microscope capable of enhancing the degree of freedom of observation while keeping its structure simple. Accordingly, a laser scanning microscope includes a light source, a spectroscopic unit guiding light from the light source to a specimen and guiding the light from the specimen to a detector, light path switching units switching a light path between the spectroscopic unit and the specimen to one among a plurality of light paths with different routes, and a plurality of light deflecting units each disposed in each of the plurality of light paths.

Abstract: A spectral image processing system and method of performing robust unmixing on measurement noise. Based on an observed spectral image acquired from a specimen and emission spectral data of each of plural materials contained in the specimen, a contribution of each of the plural materials to the observed spectral image is unmixed by a process, including an evaluating step of evaluating reliability of each component of the observed spectral image based on a predicted spectral image of the observed spectral image, and a reflecting step of reflecting a result of the evaluation in a content of the unmixing.

Abstract: A proposition of the present invention is to acquire necessary data without any chasm in an observation of a specimen containing plural kinds of substances of which exciting wavelengths are different. Accordingly, a spectral observation method of the present invention, irradiating light to the specimen containing plural kinds of substances (RFP, GFP, and so on) of which exciting wavelengths are different, and detecting spectra of light emitted from the specimen, the spectral observation method sequentially acquires spectral data of substances from the one of which exciting wavelength is long while switching a wavelength of the irradiated light among respective exciting wavelengths of the plural kinds of substances, and excludes the exciting wavelength of the substance corresponding to the spectral data to be acquired from detection wavelengths of the spectra each when acquiring the spectral data of the spectral data of the respective substances.

Abstract: A spectral image processing method is capable of reducing noise while maintaining necessary information. The spectral image processing method performs processing on a spectral image of a specimen, including a step of normalizing spectra (=spectral brightness curves) of respective pixels constituting the spectral image such that their brightness levels become equal, a step of smoothing the normalized spectra in spatial directions of the respective pixels, and a step of denormalization of multiplying spectra of the respective pixels obtained by the smoothing by either one of brightness levels of the pixels corresponding the spectra and values corresponding to the brightness levels. Consequently, the noise can be reduced while information on brightness distribution on the image is maintained.

Abstract: A confocal microscope apparatus is capable of obtaining a high confocal effect while detecting an image of a specimen line by line. The confocal microscope apparatus may include an illuminating optical system which illuminates a line-shaped area on a specimen plane in a specimen by collected light, an image-forming optical system which forms an image of light emitted from the specimen plane, a two-dimensional light detector which is placed at a conjugate plane of the specimen plane, a scanning unit which moves the line-shaped area on the specimen plane, and a correcting unit which corrects a pixel signal of a specific line on the two-dimensional light detector having a confocal relation with the line-shaped area based on a pixel signal of a peripheral line of the specific line.

Abstract: A confocal microscope apparatus capable of increasing a degree of freedom in varying sectioning resolution while keeping a configuration of a confocal microscope simple. The confocal microscope apparatus includes a confocal microscope capable of detecting two or more microscopic appearances with different sectioning and an arithmetic unit that performs arithmetic operations on data of the two or more microscopic appearances detected by the confocal microscope to create data of a microscopic appearance at sectioning resolution different from that of those microscopic appearances. Even if the number of actually measured microscopic appearances is two, it is possible to vary the diameter of a virtual-confocal diaphragm by performing arithmetic operations thereon, and therefore, an arbitrary sectioning resolution can be realized.

Abstract: It is an object to realize smooth switching between free scanning and high speed scanning in a light scanning apparatus and a light scanning microscope. To attain the object, a light scanning apparatus includes at least three mirror scanners disposed at predetermined positions of a light path for light scanning, and a light path switching unit switching the light path between a light path in which a highest-speed mirror scanner among the mirror scanners is valid and a light path in which the highest-speed mirror scanner is invalid. Therefore, the switching between a free scanning mode and a high speed scanning mode is performed by the driving of the light switching unit and involves no movement of galvanometer scanners. In addition, since the resonant galvanometer scanner is invalid during the free scanning mode, it is possible to make the resonant galvanometer scanner on standby.

Abstract: A microscope system includes a casing in which an optical element is housed, an airtight specimen chamber, and an environment control unit that controls temperature and humidity in the specimen chamber and also controls temperature inside the casing. An upper plate of the specimen chamber is an open/close lid opened and closed when inserting or removing a biological specimen. The casing becomes sealed by a bottom plate of the specimen chamber as the specimen chamber is set on the casing.